{"product_id":"highperformance-gradient-elution-the-practical-application-of-the-linearsolventstrength-model-9780471706465","title":"HighPerformance Gradient Elution The Practical","description":"\u003cb\u003eBook Synopsis\u003c\/b\u003e\u003cbr\u003eWritten by the preeminent authorities in liquid chromatography, High Performance Gradient Elution Liquid Chromatography: The Linear-Solvent-Strength Model takes the mystery out of the practice of gradient elution and helps remove barriers to practical application of this important separation technique.\u003cbr\u003e\u003cbr\u003e\u003cb\u003eTrade Review\u003c\/b\u003e\u003cbr\u003e\"This book is clear, well written, and easy to understand despite the complexity of the subject.\" (\u003ci\u003eJournal of the American Chemical Society\u003c\/i\u003e, July 2007)\u003cbr\u003e\u003cbr\u003e\u003cb\u003eTable of Contents\u003c\/b\u003e\u003cbr\u003e\u003cp\u003ePreface xv\u003c\/p\u003e \u003cp\u003eGlossary of Symbols and Terms xxi\u003c\/p\u003e \u003cp\u003e\u003cb\u003e1 Introduction 1\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e1.1 The “General Elution Problem” and the Need for Gradient Elution 1\u003c\/p\u003e \u003cp\u003e1.2 Other Reasons for the Use of Gradient Elution 4\u003c\/p\u003e \u003cp\u003e1.3 Gradient Shape 7\u003c\/p\u003e \u003cp\u003e1.4 Similarity of Isocratic and Gradient Elution 10\u003c\/p\u003e \u003cp\u003e1.4.1 Gradient and Isocratic Elution Compared 10\u003c\/p\u003e \u003cp\u003e1.4.2 The Linear-Solvent-Strength Model 13\u003c\/p\u003e \u003cp\u003e1.5 Computer Simulation 18\u003c\/p\u003e \u003cp\u003e1.6 Sample Classification 19\u003c\/p\u003e \u003cp\u003e1.6.1 Sample Compounds of Related Structure (“Regular Samples”) 19\u003c\/p\u003e \u003cp\u003e1.6.2 Sample Compounds of Unrelated Structure (“Irregular” Samples) 19\u003c\/p\u003e \u003cp\u003e\u003cb\u003e2 Gradient Elution Fundamentals 23\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e2.1 Isocratic Separation 23\u003c\/p\u003e \u003cp\u003e2.1.1 Retention 23\u003c\/p\u003e \u003cp\u003e2.1.2 Peak Width and Plate Number 24\u003c\/p\u003e \u003cp\u003e2.1.3 Resolution 25\u003c\/p\u003e \u003cp\u003e2.1.4 Role of Separation Conditions 27\u003c\/p\u003e \u003cp\u003e2.1.4.1 Optimizing Retention [Term \u003ci\u003ea\u003c\/i\u003e of Equation (2.7)] 27\u003c\/p\u003e \u003cp\u003e2.1.4.2 Optimizing Selectivity a [Term \u003ci\u003eb\u003c\/i\u003e of Equation (2.7)] 28\u003c\/p\u003e \u003cp\u003e2.1.4.3 Optimizing the Column Plate Number N [Term c of Equation (2.7)] 28\u003c\/p\u003e \u003cp\u003e2.2 Gradient Separation 31\u003c\/p\u003e \u003cp\u003e2.2.1 Retention 32\u003c\/p\u003e \u003cp\u003e2.2.1.1 Gradient and Isocratic Separation Compared for “Corresponding” Conditions 34\u003c\/p\u003e \u003cp\u003e2.2.2 Peak Width 38\u003c\/p\u003e \u003cp\u003e2.2.3 Resolution 39\u003c\/p\u003e \u003cp\u003e2.2.3.1 Resolution as a Function of Values of S for Two Adjacent Peaks (“Irregular” Samples) 42\u003c\/p\u003e \u003cp\u003e2.2.3.2 Using Gradient Elution to Predict Isocratic Separation 45\u003c\/p\u003e \u003cp\u003e2.2.4 Sample Complexity and Peak Capacity 47\u003c\/p\u003e \u003cp\u003e2.3 Effect of Gradient Conditions on Separation 49\u003c\/p\u003e \u003cp\u003e2.3.1 Gradient Steepness b: Change in Gradient Time 50\u003c\/p\u003e \u003cp\u003e2.3.2 Gradient Steepness b: Change in Column Length or Diameter 51\u003c\/p\u003e \u003cp\u003e2.3.3 Gradient Steepness b: Change in Flow Rate 55\u003c\/p\u003e \u003cp\u003e2.3.4 Gradient Range ∆\u003ci\u003eΦ\u003c\/i\u003e: Change in Initial Percentage B (\u003ci\u003eΦ\u003c\/i\u003e\u003csub\u003e0\u003c\/sub\u003e) 58\u003c\/p\u003e \u003cp\u003e2.3.5 Gradient Range ∆\u003ci\u003eΦ\u003c\/i\u003e: Change in Final Percentage B (\u003ci\u003eΦ\u003c\/i\u003e\u003csub\u003ef\u003c\/sub\u003e) 60\u003c\/p\u003e \u003cp\u003e2.3.6 Effect of a Gradient Delay 63\u003c\/p\u003e \u003cp\u003e2.3.6.1 Equipment Dwell Volume 66\u003c\/p\u003e \u003cp\u003e2.3.7 Effect of Gradient Shape (Nonlinear Gradients) 67\u003c\/p\u003e \u003cp\u003e2.3.8 Overview of the Effect of Gradient Conditions on the Chromatogram 71\u003c\/p\u003e \u003cp\u003e2.4 Related Topics 72\u003c\/p\u003e \u003cp\u003e2.4.1 Nonideal Retention in Gradient Elution 72\u003c\/p\u003e \u003cp\u003e2.4.2 Gradient Elution Misconceptions 72\u003c\/p\u003e \u003cp\u003e\u003cb\u003e3 Method Development 74\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e3.1 A Systematic Approach to Method Development 74\u003c\/p\u003e \u003cp\u003e3.1.1 Separation Goals (Step 1 of Fig. 3.1) 75\u003c\/p\u003e \u003cp\u003e3.1.2 Nature of the Sample (Step 2 of Fig. 3.1) 78\u003c\/p\u003e \u003cp\u003e3.1.3 Initial Experimental Conditions 79\u003c\/p\u003e \u003cp\u003e3.1.4 Repeatable Results 79\u003c\/p\u003e \u003cp\u003e3.1.5 Computer Simulation: Yes or No? 80\u003c\/p\u003e \u003cp\u003e3.1.6 Sample Preparation (Pretreatment) 81\u003c\/p\u003e \u003cp\u003e3.2 Initial Experiments 81\u003c\/p\u003e \u003cp\u003e3.2.1 Interpreting the Initial Chromatogram (Step 3 of Fig. 3.1) 85\u003c\/p\u003e \u003cp\u003e3.2.1.1 “Trimming” a Gradient Chromatogram 87\u003c\/p\u003e \u003cp\u003e3.2.1.2 Possible Problems 88\u003c\/p\u003e \u003cp\u003e3.3 Developing a Gradient Separation: Resolution versus Conditions 90\u003c\/p\u003e \u003cp\u003e3.3.1 Optimizing Gradient Retention k* (Step 4 of Fig. 3.1) 92\u003c\/p\u003e \u003cp\u003e3.3.2 Optimizing Gradient Selectivity a* (Step 5 of Fig. 3.1) 92\u003c\/p\u003e \u003cp\u003e3.3.3 Optimizing the Gradient Range (Step 6 of Fig. 3.1) 95\u003c\/p\u003e \u003cp\u003e3.3.3.1 Changes in Selectivity as a Result of Change in k* 96\u003c\/p\u003e \u003cp\u003e3.3.4 Segmented (Nonlinear) Gradients (Step 6 of Fig. 3.1 Continued) 100\u003c\/p\u003e \u003cp\u003e3.3.5 Optimizing the Column Plate Number N* (Step 7 of Fig. 3.1) 102\u003c\/p\u003e \u003cp\u003e3.3.6 Column Equilibration Between Successive Sample Injections 106\u003c\/p\u003e \u003cp\u003e3.3.7 Fast Separations 106\u003c\/p\u003e \u003cp\u003e3.4 Computer Simulation 108\u003c\/p\u003e \u003cp\u003e3.4.1 Quantitative Predictions and Resolution Maps 109\u003c\/p\u003e \u003cp\u003e3.4.2 Gradient Optimization 111\u003c\/p\u003e \u003cp\u003e3.4.3 Changes in Column Conditions 112\u003c\/p\u003e \u003cp\u003e3.4.4 Separation of “Regular” Samples 114\u003c\/p\u003e \u003cp\u003e3.4.5 Other Features 115\u003c\/p\u003e \u003cp\u003e3.4.5.1 Isocratic Prediction (5 in Table 3.5) 115\u003c\/p\u003e \u003cp\u003e3.4.5.2 Designated Peak Selection (6 in Table 3.5) 117\u003c\/p\u003e \u003cp\u003e3.4.5.3 Change in Other Conditions (7 in Table 3.5) 117\u003c\/p\u003e \u003cp\u003e3.4.5.4 Computer-Selection of the Best Multisegment Gradient (8 in Table 3.5) 117\u003c\/p\u003e \u003cp\u003e3.4.5.5 “Two-Run” Procedures for the Improvement of Sample Resolution 119\u003c\/p\u003e \u003cp\u003e3.4.6 Accuracy of Computer Simulation 119\u003c\/p\u003e \u003cp\u003e3.4.7 Peak Tracking 119\u003c\/p\u003e \u003cp\u003e3.5 Method Reproducibility and Related Topics 120\u003c\/p\u003e \u003cp\u003e3.5.1 Method Development 121\u003c\/p\u003e \u003cp\u003e3.5.2 Routine Analysis 122\u003c\/p\u003e \u003cp\u003e3.5.3 Change in Column Volume 123\u003c\/p\u003e \u003cp\u003e3.6 Additional Means for an Increase in Separation Selectivity 124\u003c\/p\u003e \u003cp\u003e3.7 Orthogonal Separations 127\u003c\/p\u003e \u003cp\u003e3.7.1 Two-Dimensional Separations 128\u003c\/p\u003e \u003cp\u003e4 Gradient Equipment 133\u003c\/p\u003e \u003cp\u003e4.1 Gradient System Design 133\u003c\/p\u003e \u003cp\u003e4.1.1 High-Pressure vs Low-Pressure Mixing 133\u003c\/p\u003e \u003cp\u003e4.1.2 Tradeoffs 135\u003c\/p\u003e \u003cp\u003e4.1.2.1 Dwell Volume 135\u003c\/p\u003e \u003cp\u003e4.1.2.2 Degassing 136\u003c\/p\u003e \u003cp\u003e4.1.2.3 Accuracy 137\u003c\/p\u003e \u003cp\u003e4.1.2.4 Solvent Volume Changes and Compressibility 137\u003c\/p\u003e \u003cp\u003e4.1.2.5 Flexibility 139\u003c\/p\u003e \u003cp\u003e4.1.2.6 Independent Module Use 140\u003c\/p\u003e \u003cp\u003e4.1.3 Other System Components 140\u003c\/p\u003e \u003cp\u003e4.1.3.1 Autosampler 140\u003c\/p\u003e \u003cp\u003e4.1.3.2 Column 140\u003c\/p\u003e \u003cp\u003e4.1.3.3 Detector 141\u003c\/p\u003e \u003cp\u003e4.1.3.4 Data System 141\u003c\/p\u003e \u003cp\u003e4.1.3.5 Extra-Column Volume 142\u003c\/p\u003e \u003cp\u003e4.2 General Considerations in System Selection 142\u003c\/p\u003e \u003cp\u003e4.2.1 Which Vendor? 143\u003c\/p\u003e \u003cp\u003e4.2.2 High-Pressure or Low-Pressure Mixing? 144\u003c\/p\u003e \u003cp\u003e4.2.3 Who Will Fix It? 144\u003c\/p\u003e \u003cp\u003e4.2.4 Special Applications 144\u003c\/p\u003e \u003cp\u003e4.3 Measuring Gradient System Performance 145\u003c\/p\u003e \u003cp\u003e4.3.1 Gradient Performance Test 146\u003c\/p\u003e \u003cp\u003e4.3.1.1 Gradient Linearity 146\u003c\/p\u003e \u003cp\u003e4.3.1.2 Dwell Volume Determination 147\u003c\/p\u003e \u003cp\u003e4.3.1.3 Gradient Step-Test 147\u003c\/p\u003e \u003cp\u003e4.3.1.4 Gradient Proportioning Valve Test 148\u003c\/p\u003e \u003cp\u003e4.3.2 Additional System Checks 149\u003c\/p\u003e \u003cp\u003e4.3.2.1 Flow Rate Check 149\u003c\/p\u003e \u003cp\u003e4.3.2.2 Pressure Bleed-Down 150\u003c\/p\u003e \u003cp\u003e4.3.2.3 Retention Reproducibility 150\u003c\/p\u003e \u003cp\u003e4.3.2.4 Peak Area Reproducibility 151\u003c\/p\u003e \u003cp\u003e4.4 Dwell Volume Considerations 151\u003c\/p\u003e \u003cp\u003e5 Separation Artifacts and Troubleshooting 153\u003c\/p\u003e \u003cp\u003e5.1 Avoiding Problems 154\u003c\/p\u003e \u003cp\u003e5.1.1 Equipment Checkout 157\u003c\/p\u003e \u003cp\u003e5.1.1.1 Installation Qualification, Operational Qualification, and Performance Qualification 157\u003c\/p\u003e \u003cp\u003e5.1.2 Dwell Volume 158\u003c\/p\u003e \u003cp\u003e5.1.3 Blank Gradient 158\u003c\/p\u003e \u003cp\u003e5.1.4 Suggestions for Routine Applications 158\u003c\/p\u003e \u003cp\u003e5.1.4.1 Reagent Quality 159\u003c\/p\u003e \u003cp\u003e5.1.4.2 System Cleanliness 159\u003c\/p\u003e \u003cp\u003e5.1.4.3 Degassing 159\u003c\/p\u003e \u003cp\u003e5.1.4.4 Dedicated Columns 159\u003c\/p\u003e \u003cp\u003e5.1.4.5 Equilibration 159\u003c\/p\u003e \u003cp\u003e5.1.4.6 Priming Injections 159\u003c\/p\u003e \u003cp\u003e5.1.4.7 Ignore the First Injection 160\u003c\/p\u003e \u003cp\u003e5.1.4.8 System Suitability 160\u003c\/p\u003e \u003cp\u003e5.1.4.9 Standards and Calibrators 160\u003c\/p\u003e \u003cp\u003e5.1.5 Method Development 160\u003c\/p\u003e \u003cp\u003e5.1.5.1 Use a Clean and Stable Column 160\u003c\/p\u003e \u003cp\u003e5.1.5.2 Use Reasonable Mobile Phase Conditions 161\u003c\/p\u003e \u003cp\u003e5.1.5.3 Clean Samples 162\u003c\/p\u003e \u003cp\u003e5.1.5.4 Reproducible Runs 162\u003c\/p\u003e \u003cp\u003e5.1.5.5 Sufficient Equilibration 162\u003c\/p\u003e \u003cp\u003e5.1.5.6 Reference Conditions 162\u003c\/p\u003e \u003cp\u003e5.1.5.7 Additional Tests 162\u003c\/p\u003e \u003cp\u003e5.2 Method Transfer 163\u003c\/p\u003e \u003cp\u003e5.2.1 Compensating for Dwell Volume Differences 163\u003c\/p\u003e \u003cp\u003e5.2.1.1 Injection Delay 163\u003c\/p\u003e \u003cp\u003e5.2.1.2 Adjustment of the Initial Isocratic Hold 164\u003c\/p\u003e \u003cp\u003e5.2.1.3 Use of Maximum-Dwell-Volume Methods 165\u003c\/p\u003e \u003cp\u003e5.2.1.4 Adjustment of Initial Percentage B 165\u003c\/p\u003e \u003cp\u003e5.2.2 Other Sources of Method Transfer Problems 168\u003c\/p\u003e \u003cp\u003e5.2.2.1 Gradient Shape 169\u003c\/p\u003e \u003cp\u003e5.2.2.2 Gradient Rounding 169\u003c\/p\u003e \u003cp\u003e5.2.2.3 Inter-Run Equilibration 169\u003c\/p\u003e \u003cp\u003e5.2.2.4 Column Size 169\u003c\/p\u003e \u003cp\u003e5.2.2.5 Column Temperature 169\u003c\/p\u003e \u003cp\u003e5.2.2.6 Interpretation of Method Instructions 170\u003c\/p\u003e \u003cp\u003e5.3 Column Equilibration 170\u003c\/p\u003e \u003cp\u003e5.3.1 Primary Effects 171\u003c\/p\u003e \u003cp\u003e5.3.2 Slow Equilibration of Column and Mobile Phase 173\u003c\/p\u003e \u003cp\u003e5.3.3 Practical Considerations and Recommendations 174\u003c\/p\u003e \u003cp\u003e5.4 Separation Artifacts 175\u003c\/p\u003e \u003cp\u003e5.4.1 Baseline Drift 176\u003c\/p\u003e \u003cp\u003e5.4.2 Baseline Noise 179\u003c\/p\u003e \u003cp\u003e5.4.2.1 Baseline Noise: A Case Study 180\u003c\/p\u003e \u003cp\u003e5.4.3 Peaks in a Blank Gradient 182\u003c\/p\u003e \u003cp\u003e5.4.3.1 Mobile Phase Water or Organic Solvent Impurities 182\u003c\/p\u003e \u003cp\u003e5.4.3.2 Other Sources of Background Peaks 185\u003c\/p\u003e \u003cp\u003e5.4.4 Extra Peaks for Injected Samples 185\u003c\/p\u003e \u003cp\u003e5.4.4.1 t\u003csub\u003e0\u003c\/sub\u003e Peaks 185\u003c\/p\u003e \u003cp\u003e5.4.4.2 Air Peaks 186\u003c\/p\u003e \u003cp\u003e5.4.4.3 Late Peaks 187\u003c\/p\u003e \u003cp\u003e5.4.5 Peak Shape Problems 188\u003c\/p\u003e \u003cp\u003e5.4.5.1 Tailing and Fronting 188\u003c\/p\u003e \u003cp\u003e5.4.5.2 Excess Peak Broadening 188\u003c\/p\u003e \u003cp\u003e5.4.5.3 Split Peaks 190\u003c\/p\u003e \u003cp\u003e5.4.5.4 Injection Conditions 191\u003c\/p\u003e \u003cp\u003e5.4.5.5 Sample Decomposition 193\u003c\/p\u003e \u003cp\u003e5.5 Troubleshooting 195\u003c\/p\u003e \u003cp\u003e5.5.1 Problem Isolation 196\u003c\/p\u003e \u003cp\u003e5.5.2 Troubleshooting and Maintenance Suggestions 197\u003c\/p\u003e \u003cp\u003e5.5.2.1 Removing Air from the Pump 197\u003c\/p\u003e \u003cp\u003e5.5.2.2 Solvent Siphon Test 197\u003c\/p\u003e \u003cp\u003e5.5.2.3 Premixing to Improve Retention Reproducibility in Shallow Gradients 198\u003c\/p\u003e \u003cp\u003e5.5.2.4 Cleaning and Handling Check-Valves 199\u003c\/p\u003e \u003cp\u003e5.5.2.5 Replacing Pump Seals and Pistons 200\u003c\/p\u003e \u003cp\u003e5.5.2.6 Leak Detection 200\u003c\/p\u003e \u003cp\u003e5.5.2.7 Repairing Fitting Leaks 200\u003c\/p\u003e \u003cp\u003e5.5.2.8 Cleaning Glassware 201\u003c\/p\u003e \u003cp\u003e5.5.2.9 For Best Results with TFA 201\u003c\/p\u003e \u003cp\u003e5.5.2.10 Improved Water Purity 201\u003c\/p\u003e \u003cp\u003e5.5.2.11 Isolating Carryover Problems 203\u003c\/p\u003e \u003cp\u003e5.5.2.12 Troubleshooting Rules of Thumb 204\u003c\/p\u003e \u003cp\u003e5.5.3 Gradient Performance Test Failures 206\u003c\/p\u003e \u003cp\u003e5.5.3.1 Linearity (4.3.1.1) 206\u003c\/p\u003e \u003cp\u003e5.5.3.2 Step Test (4.3.1.3) 206\u003c\/p\u003e \u003cp\u003e5.5.3.3 Gradient-Proportioning-Valve Test (4.3.1.4) 209\u003c\/p\u003e \u003cp\u003e5.5.3.4 Flow Rate (4.3.2.1) 211\u003c\/p\u003e \u003cp\u003e5.5.3.5 Pressure Bleed-Down (4.3.2.2) 212\u003c\/p\u003e \u003cp\u003e5.5.3.6 Retention Reproducibility (4.3.2.3) 212\u003c\/p\u003e \u003cp\u003e5.5.3.7 Peak Area Reproducibility (4.3.2.4) 213\u003c\/p\u003e \u003cp\u003e5.5.4 Troubleshooting Case Studies 213\u003c\/p\u003e \u003cp\u003e5.5.4.1 Retention Variation – Case Study 1 213\u003c\/p\u003e \u003cp\u003e5.5.4.2 Retention Variation – Case Study 2 218\u003c\/p\u003e \u003cp\u003e5.5.4.3 Contaminated Reagents – Case Study 3 220\u003c\/p\u003e \u003cp\u003e5.5.4.4 Baseline and Retention Problems – Case Study 4 224\u003c\/p\u003e \u003cp\u003e6 Separation of Large Molecules 228\u003c\/p\u003e \u003cp\u003e6.1 General Considerations 228\u003c\/p\u003e \u003cp\u003e6.1.1 Values of S for Large Molecules 229\u003c\/p\u003e \u003cp\u003e6.1.2 Values of N* for Large Molecules 235\u003c\/p\u003e \u003cp\u003e6.1.3 Conformational State 236\u003c\/p\u003e \u003cp\u003e6.1.4 Homo-Oligomeric Samples 238\u003c\/p\u003e \u003cp\u003e6.1.4.1 Separation of Large Homopolymers 241\u003c\/p\u003e \u003cp\u003e6.1.5 Proposed Models for the Gradient Separation of Large Molecules 242\u003c\/p\u003e \u003cp\u003e6.1.5.2 “Critical Elution Behavior”: Biopolymers 246\u003c\/p\u003e \u003cp\u003e6.1.5.3 Measurement of LSS Parameters for Large Molecules 247\u003c\/p\u003e \u003cp\u003e6.2 Biomolecules 248\u003c\/p\u003e \u003cp\u003e6.2.1 Peptides and Proteins 248\u003c\/p\u003e \u003cp\u003e6.2.1.1 Sample Characteristics 249\u003c\/p\u003e \u003cp\u003e6.2.1.2 Conditions for an Initial Gradient Run 249\u003c\/p\u003e \u003cp\u003e6.2.1.3 Method Development 253\u003c\/p\u003e \u003cp\u003e6.2.1.4 Segmented Gradients 259\u003c\/p\u003e \u003cp\u003e6.2.2 Other Separation Modes and Samples 261\u003c\/p\u003e \u003cp\u003e6.2.2.1 Hydrophobic Interaction Chromatography 262\u003c\/p\u003e \u003cp\u003e6.2.2.2 Ion Exchange Chromatography 264\u003c\/p\u003e \u003cp\u003e6.2.2.3 Hydrophilic Interaction Chromatography 266\u003c\/p\u003e \u003cp\u003e6.2.2.4 Separation of Viruses 267\u003c\/p\u003e \u003cp\u003e6.2.3 Separation Problems 271\u003c\/p\u003e \u003cp\u003e6.2.4 Fast Separations of Peptides and Proteins 274\u003c\/p\u003e \u003cp\u003e6.2.5 Two-Dimensional Separations of Peptides and Proteins 274\u003c\/p\u003e \u003cp\u003e6.3 Synthetic Polymers 275\u003c\/p\u003e \u003cp\u003e6.3.1 Determination of Molecular Weight Distribution 277\u003c\/p\u003e \u003cp\u003e6.3.2 Determination of Chemical Composition 278\u003c\/p\u003e \u003cp\u003e7 Preparative Separations 283\u003c\/p\u003e \u003cp\u003e7.1 Introduction 283\u003c\/p\u003e \u003cp\u003e7.1.1 Equipment for Preparative Separation 285\u003c\/p\u003e \u003cp\u003e7.2 Isocratic Separation 286\u003c\/p\u003e \u003cp\u003e7.2.1 Touching-Peak Separation 287\u003c\/p\u003e \u003cp\u003e7.2.1.1 Theory 287\u003c\/p\u003e \u003cp\u003e7.2.1.2 Column Saturation Capacity 289\u003c\/p\u003e \u003cp\u003e7.2.1.3 Sample-Volume Overload 292\u003c\/p\u003e \u003cp\u003e7.2.2 Method Development for Isocratic Touching-Peak Separation 292\u003c\/p\u003e \u003cp\u003e7.2.2.1 Optimizing Separation Conditions 295\u003c\/p\u003e \u003cp\u003e7.2.2.2 Selecting a Sample Weight for Touching-Peak Separation 297\u003c\/p\u003e \u003cp\u003e7.2.2.3 Scale-Up 298\u003c\/p\u003e \u003cp\u003e7.2.2.4 Sample Solubility 300\u003c\/p\u003e \u003cp\u003e7.2.3 Beyond Touching-Peak Separation 301\u003c\/p\u003e \u003cp\u003e7.3 Gradient Separation 302\u003c\/p\u003e \u003cp\u003e7.3.1 Touching-Peak Separation 306\u003c\/p\u003e \u003cp\u003e7.3.2 Method Development for Gradient Touching-Peak Separation 306\u003c\/p\u003e \u003cp\u003e7.3.2.1 Step Gradients 311\u003c\/p\u003e \u003cp\u003e7.3.3 Sample-Volume Overload 312\u003c\/p\u003e \u003cp\u003e7.3.4 Possible Complications of Simple Touching-Peak Theory and Their Practical Impact 312\u003c\/p\u003e \u003cp\u003e7.3.4.1 Crossing Isotherms 313\u003c\/p\u003e \u003cp\u003e7.3.4.2 Unequal Values of S 314\u003c\/p\u003e \u003cp\u003e7.4 Severely Overloaded Separation 315\u003c\/p\u003e \u003cp\u003e7.4.1 Is Gradient Elution Necessary? 316\u003c\/p\u003e \u003cp\u003e7.4.2 Displacement Effects 317\u003c\/p\u003e \u003cp\u003e7.4.3 Method Development 317\u003c\/p\u003e \u003cp\u003e7.4.4 Separations of Peptides and Small Proteins 318\u003c\/p\u003e \u003cp\u003e7.4.5 Column Efficiency 320\u003c\/p\u003e \u003cp\u003e7.4.6 Production-Scale Separation 320\u003c\/p\u003e \u003cp\u003e8 other Applications of Gradient Elution 323\u003c\/p\u003e \u003cp\u003e8.1 Gradient Elution for LC-MS 324\u003c\/p\u003e \u003cp\u003e8.1.1 Application Areas 325\u003c\/p\u003e \u003cp\u003e8.1.2 Requirements for LC-MS 325\u003c\/p\u003e \u003cp\u003e8.1.3 Basic LC-MS Concepts 326\u003c\/p\u003e \u003cp\u003e8.1.3.1 The Interface 326\u003c\/p\u003e \u003cp\u003e8.1.3.2 Column Configurations 328\u003c\/p\u003e \u003cp\u003e8.1.3.3 Quadrupoles and Ion Traps 328\u003c\/p\u003e \u003cp\u003e8.1.4 LC-UV vs LC-MS Gradient Conditions 330\u003c\/p\u003e \u003cp\u003e8.1.5 Method Development for LC-MS 332\u003c\/p\u003e \u003cp\u003e8.1.5.1 Define Separation Goals (Step 1, Table 8.2) 332\u003c\/p\u003e \u003cp\u003e8.1.5.2 Collect Information on Sample (Step 2, Table 8.2) 334\u003c\/p\u003e \u003cp\u003e8.1.5.3 Carry Out Initial Separation (Run 1, Step 3, Table 8.2) 339\u003c\/p\u003e \u003cp\u003e8.1.5.4 Optimize Gradient Retention k* (Step 4, Table 8.2) 339\u003c\/p\u003e \u003cp\u003e8.1.5.5 Optimize Selectivity a* (Step 5, Table 8.2) 339\u003c\/p\u003e \u003cp\u003e8.1.5.6 Adjust Gradient Range and Shape (Step 6, Table 8.2) 340\u003c\/p\u003e \u003cp\u003e8.1.5.7 Vary Column Conditions (Step 7, Table 8.2) 341\u003c\/p\u003e \u003cp\u003e8.1.5.8 Determine Inter-Run Column Equilibration (Step 8, Table 8.2) 341\u003c\/p\u003e \u003cp\u003e8.1.6 Special Challenges for LC-MS 341\u003c\/p\u003e \u003cp\u003e8.1.6.1 Dwell Volume 342\u003c\/p\u003e \u003cp\u003e8.1.6.2 Gradient Distortion 342\u003c\/p\u003e \u003cp\u003e8.1.6.3 Ion Suppression 343\u003c\/p\u003e \u003cp\u003e8.1.6.4 Co-Eluting Compounds 345\u003c\/p\u003e \u003cp\u003e8.1.6.5 Resolution Requirements 346\u003c\/p\u003e \u003cp\u003e8.1.6.6 Use of Computer Simulation Software 347\u003c\/p\u003e \u003cp\u003e8.1.6.7 Isocratic Methods 347\u003c\/p\u003e \u003cp\u003e8.1.6.8 Throughput Enhancement 347\u003c\/p\u003e \u003cp\u003e8.2 Ion-Exchange Chromatography 349\u003c\/p\u003e \u003cp\u003e8.2.1 Theory 349\u003c\/p\u003e \u003cp\u003e8.2.2 Dependence of Separation on Gradient Conditions 356\u003c\/p\u003e \u003cp\u003e8.2.3 Method Development for Gradient IEC 356\u003c\/p\u003e \u003cp\u003e8.2.3.1 Choice of Initial Conditions 356\u003c\/p\u003e \u003cp\u003e8.2.3.2 Improving the Separation 357\u003c\/p\u003e \u003cp\u003e8.3 Normal-Phase Chromatography 359\u003c\/p\u003e \u003cp\u003e8.3.1 Theory 359\u003c\/p\u003e \u003cp\u003e8.3.2 Method Development for Gradient NPC 360\u003c\/p\u003e \u003cp\u003e8.3.3 Hydrophilic Interaction Chromatography 361\u003c\/p\u003e \u003cp\u003e8.3.3.1 Method Development for Gradient HILIC 361\u003c\/p\u003e \u003cp\u003e8.4 Ternary- or Quaternary-Solvent Gradients 365\u003c\/p\u003e \u003cp\u003e9 Theory and Derivations 370\u003c\/p\u003e \u003cp\u003e9.1 The Linear Solvent Strength Model 370\u003c\/p\u003e \u003cp\u003e9.1.1 Retention 372\u003c\/p\u003e \u003cp\u003e9.1.1.1 Gradient and Isocratic Retention Compared 374\u003c\/p\u003e \u003cp\u003e9.1.1.2 Small Values of k \u003csub\u003e0\u003c\/sub\u003e 376\u003c\/p\u003e \u003cp\u003e9.1.2 Peak Width 378\u003c\/p\u003e \u003cp\u003e9.1.2.1 Gradient Compression 380\u003c\/p\u003e \u003cp\u003e9.1.3 Selectivity and Resolution 383\u003c\/p\u003e \u003cp\u003e9.1.4 Advantages of LSS Behavior 385\u003c\/p\u003e \u003cp\u003e9.2 Second-Order Effects 386\u003c\/p\u003e \u003cp\u003e9.2.1 Assumptions About Φ and k 386\u003c\/p\u003e \u003cp\u003e9.2.1.1 Incomplete Column Equilibration 386\u003c\/p\u003e \u003cp\u003e9.2.1.2 Solvent Demixing 391\u003c\/p\u003e \u003cp\u003e9.2.1.3 Nonlinear Plots of log k vs Φ 393\u003c\/p\u003e \u003cp\u003e9.2.1.4 Dependence of V \u003csub\u003em\u003c\/sub\u003e on Φ 393\u003c\/p\u003e \u003cp\u003e9.2.2 Nonideal Equipment 393\u003c\/p\u003e \u003cp\u003e9.3. Accuracy of Gradient Elution Predictions 397\u003c\/p\u003e \u003cp\u003e9.3.1 Gradient Retention Time 397\u003c\/p\u003e \u003cp\u003e9.3.1.1 Confirmation of Equation (9.2) 397\u003c\/p\u003e \u003cp\u003e9.3.1.2 Computer Simulation 399\u003c\/p\u003e \u003cp\u003e9.3.2 Peak Width Predictions 399\u003c\/p\u003e \u003cp\u003e9.3.3 Measurement of Values of S and log k \u003csub\u003e0\u003c\/sub\u003e 400\u003c\/p\u003e \u003cp\u003e9.4 Values of S 401\u003c\/p\u003e \u003cp\u003e9.4.1 Estimating Values of S from Solute Properties and Experimental Conditions 402\u003c\/p\u003e \u003cp\u003e9.5 Values of N in Gradient Elution 404\u003c\/p\u003e \u003cp\u003eAppendix I The Constant-S Approximation In Gradient Elution 414\u003c\/p\u003e \u003cp\u003eAppendix II Estimation of Conditions for Isocratic Elution, Based on An Initial Gradient Run 416\u003c\/p\u003e \u003cp\u003eAppendix III Characterization of Reversed-phase Columns for Selectivity and Peak Tailing 418\u003c\/p\u003e \u003cp\u003eAppendix IV Solvent Properties Relevant to the Use of Gradient Elution 434\u003c\/p\u003e \u003cp\u003eAppendix V Theory Of Preparative Separation 436\u003c\/p\u003e \u003cp\u003eAppendix Vi Further Information On Virus Chromatography 445\u003c\/p\u003e \u003cp\u003eIndex 450\u003c\/p\u003e","brand":"John Wiley \u0026 Sons Inc","offers":[{"title":"Default Title","offer_id":49402656817495,"sku":"9780471706465","price":138.56,"currency_code":"GBP","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0817\/1739\/5799\/files\/9780471706465.jpg?v=1730481139","url":"https:\/\/bookcurl.com\/products\/highperformance-gradient-elution-the-practical-application-of-the-linearsolventstrength-model-9780471706465","provider":"Book Curl","version":"1.0","type":"link"}